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Antimicrob. Agents Chemother. doi:10.1128/AAC.01569-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Isolation and Purification of Enterocin E-760 with Broad Antimicrobial Activity Against Gram-positive and Gram-negative Bacteria

USDA-ARS, Russell Research Center, Poultry Microbiological Safety Research Unit Athens, GA, U.S.A.; State Research Center for Applied Microbiology and Biotechnology, Obolensk, Russian Federation

^* To whom correspondence should be addressed. Email: eric.line{at}ars.usda.gov.

	Abstract

Strain NRRL B-30745, isolated from chicken ceca, and identified as Enterococcus durans/faecium/hirae was initially identified as antagonistic to Campylobacter jejuni. The isolate produced a 5362 Da bacteriocin (enterocin) that inhibits the growth of Salmonella enteritidis, S. choleraesuis, S. typhimurium, S. gallinarum, Escherichia coli O157:H7, Yersinia enterocolitica, Citrobacter freundii, Klebsiella pneumoniae, Shigella dysenteriae, Pseudomonas aeruginosa, Proteus mirabilis, Morganella morganii, Staphylococcus aureus, S. epidermidis, Listeria monocytogenes, Campylobacter jejuni and 20 other Campylobacter spp. isolates. The enterocin, E-760, was isolated and purified by cation exchange and hydrophobic interaction chromatography. The proteinaceous nature of purified enterocin E-760 was demonstrated upon treatment with various proteolytic enzymes. Specifically, the antimicrobial peptide was found to be sensitive to Beta-chymotrypsin, proteinase K and papain, while it was resistant to lysozyme and lipase. The enterocin demonstrated thermostability by retaining activity after 5 min at 100°C and was stable at pH values between 5.0 and 8.7. However, activity was lost below pH 3.0 and above pH 9.5. Administration of enterocin E-760 treated feed significantly (P< 0.05) reduced colonization of young broiler chicks experimentally challenged and colonized with two strains of C. jejuni by more than 8 log₁₀ CFU. Enterocin E-760 also significantly (P<0.05) reduced colonization of naturally acquired Campylobacter spp. in market age broilers when administered in treated feed four days prior to analysis.